Quantification of matrix metalloproteinases and tissue inhibitors of metalloproteinase in prostatic tissue: analytical aspects.
BACKGROUND
The balance between matrix metalloproteinases (MMP) and the tissue inhibitors of metalloproteinases (TIMP) has been seen as important during tumor invasion and progression. The determination of these components needs a special strategy of tissue preparation. This analytical problem has not been considered for prostatic tissue.


METHODS
We adapted an extraction method consisting of two extraction steps with 0.25% Triton X-100/CaCl2 solution and two heat extraction steps at 60 degrees C for 4 min. This combination allowed a complete extraction of MMP (measured as enzyme activity) and TIMP-1 (measured with an ELISA test) from cancerous and normal prostatic tissue samples.


RESULTS
The median values for cancerous vs. normal MMPs (50.8 mU/g wet tissue and 1,580 mU/g protein vs. 88.8 and 2,497) and TIMP-1 (4.49 micrograms/g wet tissue and 96.7 micrograms/g protein vs. 12.4 and 237.8) were significantly lower, whereas the respective ratios for MMP/ TIMP-1 (11.1 vs. 4.0 on wet weight and 15.5 vs. 5.3 on protein basis) were significantly higher.


CONCLUSIONS
An optimized extraction procedure was elaborated for determining MMPs and TIMP-1 in prostatic tissue samples. The increased ratio of MMP/TIMP-1 can be interpreted as an indicator of the imbalance between MMP and TIMP, characteristic of prostate carcinoma tissue.
Jung
K
K
Department of Urology, University Hospital Charité, Humboldt University, Berlin, Germany. jung@rz.charite.hu-berlin.de
Lein
M
M
Ulbrich
N
N
Rudolph
B
B
Henke
W
W
Schnorr
D
D
Loening
S A
SA
D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D008297:Male; D008666:Metalloendopeptidases / Q000032:analysis*; D008875:Middle Aged; D011467:Prostate / Q000737:chemistry* / Q000166:cytology; D011471:Prostatic Neoplasms / Q000737:chemistry* / Q000473:pathology; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000032:analysis; D019714:Tissue Inhibitor of Metalloproteinases / Q000032:analysis*
D019715:Tissue Inhibitor of Metalloproteinase-1; D019714:Tissue Inhibitor of Metalloproteinases; D008666:Metalloendopeptidases